Quantitative measurement of cysteinyl leukotrienes and leukotriene B4 in human sputum using ultra high pressure liquid chromatography–tandem mass spectrometry

摘要: Abstract The role of leukotrienes (LTs) in airway inflammatory diseases, such as asthma, has been extensively reported. measurement LTs sputum supernatants, which is commonly done via enzyme immunoassays (EIAs), may prove to be useful for assessing inflammation. Despite the many advantages EIA, these methods suffer from a lack selectivity. Therefore, selective and reliable method analysis human needed. In this study we developed validated sensitive specific using ultra high pressure liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS), measure simultaneously cysteinyl (CysLTs) leukotriene B4 (LTB4) sputum. Sputum supernatants obtained by ultracentrifugation were stabilized protease inhibitors, spiked with stable isotopic internal standards, subjected solid phase extraction (SPE) UHPLC separation. Multiple reaction monitoring (MRM) transitions optimized measured on spectrometer. limit detection (LOD) LTE4 LTB4 was 9.8 19.5 pg/mL, respectively. lower quantitation (LLOQ) 19.5 39.0 pg/mL, dynamic range assay 5000 pg/mL, whereas 10,000 pg/mL. intra- inter-day % coefficient variation (%CV)